Crystalline forms of nicotinamide riboside chloride

ABSTRACT

Provided herein are crystalline forms of nicotinamide riboside chloride and methods of making the same. Also provided are compositions comprising the crystalline form of nicotinamide riboside chloride, and therapeutic methods employing the crystalline form of nicotinamide riboside chloride.

RELATED APPLICATION

This application claims the benefit of priority to U.S. ProvisionalPatent Application Ser. No. 62/609,512, filed Dec. 22, 2017, whichapplication is hereby incorporated by reference in its entirety.

BACKGROUND OF THE INVENTION

Nicotinamide riboside is a pyridine-nucleoside form of niacin (i.e.,vitamin B3) that serves as a precursor to nicotinamide adeninedinucleotide (NAD⁺). NAD⁺ promotes cellular metabolism, mitochondrialfunction, and energy production. Currently, nicotinamide riboside ismade through synthetic methods or fermentation processes. Because of itssignificant potential to confer health benefits when used as a dietarysupplement, there exists a need to develop highly efficient and scalableprocesses for the manufacture and purification of nicotinamide riboside.

SUMMARY OF THE INVENTION

In certain aspects, the present invention provides a crystalline form ofa compound having the structure of formula (I):

wherein the crystalline form is characterized by 2θ values of 20.9±0.1,21.6±0.1, 21.8±0.1, 23.6±0.1, and 24.4±0.1.

In certain aspects, the present invention provides a pharmaceuticalcomposition comprising a crystalline form of the invention incombination with a pharmaceutically acceptable carrier.

In some aspects, the invention provides a method for preparing acrystalline form of the invention, the method comprising the steps of(a) providing a mixture of a compound of formula (I) in a first organicsolvent; and (b) crystallizing the compound of formula (I) from themixture of a compound of formula (I) in a first organic solvent.

In certain aspects, the present invention provides a method of improvingcellular health in a subject, comprising administering to the subject atherapeutically effective amount of a crystalline form of the invention

In certain aspects, the present invention provides a method of improvingsleep quality, stimulating or increasing REM sleep, or treating orpreventing insomnia, desynchronosis, or a circadian rhythm sleepdisorder in a subject, comprising administering to the subject atherapeutically effective amount of a crystalline form of the invention.

In certain aspects, the present invention provides a method of treatingor preventing a motor neuron disease or ALS, or slowing or reversing theprogression of motor neuron degeneration in a subject, comprisingadministering to the subject a therapeutically effective amount of acrystalline form of the invention.

In certain aspects, the present invention provides a method of improvingfertility, treating or preventing infertility, inducing ovulation,increasing sperm count, or increasing lactation, comprisingadministering to the subject a therapeutically effective amount of acrystalline form of the invention.

In certain aspects, the present invention provides a method of treatingor preventing kidney damage, acute kidney injury, or kidney disease, orincreasing blood flow to the kidneys, comprising administering to thesubject a therapeutically effective amount of a crystalline form of theinvention.

In certain aspects, the present invention provides a method of treatingor preventing liver damage or fatty liver, or decreasing the serum levelof alanine transaminase (ALT) or aspartate transaminase (AST) in asubject, comprising administering to the subject a therapeuticallyeffective amount of a crystalline form of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an x-ray diffraction spectrum of the crystallinenicotinamide riboside chloride.

FIG. 2 shows a DSC thermogram of the crystalline nicotinamide ribosidechloride at a heating rate of 5° C./min.

FIG. 3 shows a DSC thermogram of the crystalline nicotinamide ribosidechloride at a heating rate of 2° C./min.

FIG. 4 shows a DSC thermogram of the crystalline nicotinamide ribosidechloride at a heating rate of 10° C./min.

FIG. 5 shows a DSC thermogram of the crystalline nicotinamide ribosidechloride at a heating rate of 20° C./min.

FIG. 6 shows a process flow diagram for the manufacture of thecrystalline form of nicotinamide riboside chloride.

DETAILED DESCRIPTION OF THE INVENTION I. Definitions

As used herein, the term “administering” means providing apharmaceutical agent or composition to a subject, and includes, but isnot limited to, administering by a medical professional andself-administering. Administration of a substance, a compound or anagent to a subject can be carried out using one of a variety of methodsknown to those skilled in the art. For example, a compound or an agentcan be administered, intravenously, arterially, intradermally,intramuscularly, intraperitoneally, subcutaneously, ocularly,sublingually, orally (by ingestion), intranasally (by inhalation),intraspinally, intracerebrally, and transdermally (by absorption, e.g.,through a skin duct). A compound or agent can also appropriately beintroduced by rechargeable or biodegradable polymeric devices or otherdevices, e.g., patches and pumps, or formulations, which provide for theextended, slow or controlled release of the compound or agent.Administering can also be performed, for example, once, a plurality oftimes, and/or over one or more extended periods.

Appropriate methods of administering a substance, a compound or an agentto a subject will also depend, for example, on the age and/or thephysical condition of the subject and the chemical and biologicalproperties of the compound or agent (e.g., solubility, digestibility,bioavailability, stability and toxicity). In some embodiments, acompound or an agent is administered orally, e.g., to a subject byingestion. In some embodiments, the orally administered compound oragent is in an extended release or slow release formulation, oradministered using a device for such slow or extended release.

The phrase “pharmaceutically-acceptable carrier” as used herein means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, or solvent encapsulatingmaterial.

As used herein, the term “subject” means a human or non-human animalselected for treatment or therapy.

The phrases “therapeutically-effective amount” and “effective amount” asused herein means the amount of an agent which is effective forproducing the desired therapeutic effect in at least a sub-population ofcells in a subject at a reasonable benefit/risk ratio applicable to anymedical treatment.

“Treating” a disease in a subject or “treating” a subject having adisease refers to subjecting the subject to a pharmaceutical treatment,e.g., the administration of a drug, such that at least one symptom ofthe disease is decreased or prevented from worsening.

As used herein, a therapeutic that “prevents” a disorder or conditionrefers to a compound that, when administered to a statistical sampleprior to the onset of the disorder or condition, reduces the occurrenceof the disorder or condition in the treated sample relative to anuntreated control sample, or delays the onset or reduces the severity ofone or more symptoms of the disorder or condition relative to theuntreated control sample.

II. Crystal Forms

The present invention is based on the surprising discovery of a newcrystalline form of nicotinamide riboside chloride having excellentpurity and ease of manufacture. Accordingly, in certain embodiments, theinvention provides a crystalline form of a compound having the structureof formula (I):

wherein the crystalline form is characterized by 2θ values of 20.9±0.1,21.6±0.1, 21.8±0.1, 23.6±0.1, and 24.4±0.1.

In further embodiments, the crystalline form of the invention ischaracterized by 20 values of 9.9±0.1, 18.4±0.1, 20.9±0.1, 21.6±0.1,21.8±0.1, 23.6±0.1, 24.4±0.1, 29.4±0.1, 29.9±0.1, 30.5±0.1, and33.5±0.1.

In yet further embodiments, the crystalline form of the invention ischaracterized by 20 values of 9.9±0.1, 18.4±0.1, 20.9±0.1, 21.6±0.1,21.8±0.1, 22.9±0.1, 23.6±0.1, 24.4±0.1, 25.2±0.1, 29.4±0.1, 29.9±0.1,30.5±0.1, 31.9±0.1, 32.1±0.1, 33.5±0.1, 34.1±0.1, and 37.4±0.1.

In still yet further embodiments, the crystalline form of the inventionis characterized by 2θ values of 9.9±0.1, 15.6±0.1, 18.4±0.1, 18.5±0.1,19.0±0.1, 20.9±0.1, 21.6±0.1, 21.8±0.1, 22.9±0.1, 23.6±0.1, 24.4±0.1,25.2±0.1, 29.4±0.1, 29.9±0.1, 30.5±0.1, 31.5±0.1, 31.9±0.1, 32.1±0.1,33.5±0.1, 34.0±0.1, 34.1±0.1, 36.5±0.1, and 37.4±0.1.

In some embodiments, the crystalline form of the invention ischaracterized by 2θ values of 20.87±0.10, 21.55±0.10, 21.79±0.10,23.63±0.10, and 24.44±0.10.

In further such embodiments, the crystalline form of the invention ischaracterized by 2θ values of 9.87±0.10, 18.36±0.10, 20.87±0.10,21.55±0.10, 21.79±0.10, 23.63±0.10, 24.44±0.10, 29.35±0.10, 29.93±0.10,30.47±0.10, and 33.51±0.10.

In yet further such embodiments, the crystalline form of the inventionis characterized by 2θ values of 9.87±0.10, 18.36±0.10, 20.87±0.10,21.55±0.10, 21.79±0.10, 22.87±0.10, 23.63±0.10, 24.44±0.10, 25.25±0.10,29.35±0.10, 29.93±0.10, 30.47±0.10, 31.87±0.10, 32.08±0.10, 33.51±0.10,34.15±0.10, and 37.38±0.10.

In still yet further embodiments, the crystalline form of the inventionis characterized by 2θ values 9.87±0.10, 15.61±0.10, 18.36±0.10,18.49±0.10, 19.01±0.10, 20.87±0.10, 21.55±0.10, 21.79±0.10, 22.87±0.10,23.63±0.10, 24.44±0.10, 25.25±0.10, 29.35±0.10, 29.93±0.10, 30.47±0.10,31.46±0.10, 31.87±0.10, 32.08±0.10, 33.51±0.10, 34.00±0.10, 34.15±0.10,36.53±0.10, and 37.38±0.10.

The crystalline form of the invention may have an XRD spectrumsubstantially as shown in FIG. 1.

In some embodiments, the crystalline form of the invention has anendotherm of 123.9° C. in the DSC thermogram at a heating rate of 5°C./min. For example, the crystalline form of may have a DSC thermogramsubstantially as shown in FIG. 2.

In other embodiments, the crystalline form of the invention has anendotherm of 115.4° C. in the DSC thermogram at a heating rate of 2°C./min. For example, the crystalline form of may have a DSC thermogramsubstantially as shown in FIG. 3.

In other embodiments, the crystalline form of the invention has anendotherm of 131.2° C. in the DSC thermogram at a heating rate of 10°C./min. Such a crystal form may have a DSC thermogram substantially asshown in FIG. 4.

In some embodiments, the crystalline form of the invention has anendotherm of 139.2° C. in the DSC thermogram at a heating rate of 20°C./min. Such a crystal form may have a DSC thermogram substantially asshown in FIG. 5.

In certain such embodiments, the endotherm represents the melting point.

The crystalline form of the invention may have a purity of 95%, 96%,97%, 98%, 99%, 99.2%, 99.4%, 99.6%, 99.8% or higher.

III. Synthetic Methods

In certain aspects, the present invention provides a method forpreparing a crystalline form described herein, the method comprising (a)providing a mixture of a compound of formula (I) in a first organicsolvent; and (b) crystallizing the compound of formula (I) from themixture of a compound of formula (I) in a first organic solvent.

In certain embodiments, the first organic solvent is methanol.

In alternative embodiments, the first organic solvent is another polarprotic solvent such as ethanol or isopropanol. Alternatively, the firstorganic solvent is a polar aprotic solvent, such as tetrahydrofuran,optionally in combination with water or an organic polar protic solvent.

In certain embodiments, the mixture comprising the compound of formula(I) and the first organic solvent is a solution, and the step ofcrystallizing the compound of formula (I) from the mixture comprisesbringing the solution to supersaturation to cause the compound offormula (I) to precipitate out of solution.

In certain embodiments, the step of bringing the solution tosupersaturation comprises slowly adding an anti-solvent, allowing thesolution to cool, reducing the volume of the solution, or anycombination thereof. In some embodiments, the step of bringing thesolution to supersaturation comprises cooling the solution to ambienttemperature or lower. Preferably, the step of bringing the solution tosupersaturation comprises reducing the volume of the solution.

In certain embodiments, the method further comprises isolating thecrystalline form, e.g. by filtering the crystals, by decanting fluidfrom the crystals, or by any other suitable separation technique.

In further embodiments, the preparation method further comprises washingthe crystalline form with a second organic solvent. The second organicsolvent may be selected from ethanol, acetone, methyl tert-butyl etherand combinations thereof.

The method can also comprise the step of drying the crystals, forexample under reduced pressure.

A “polar protic solvent” as used herein is a solvent having a dipolemoment of about 1.4 to 4.0 D, and comprising a chemical moiety thatparticipates in hydrogen bonding, such as an O—H bond or an N—H bond.Exemplary polar protic solvents include methanol, ethanol, n-propanol,isopropanol, n-butanol, isobutanol, ammonia, water, and acetic acid.

A “polar aprotic solvent” as used herein means a solvent having a dipolemoment of about 1.4 to 4.0 D that lacks a hydrogen bonding group such asO—H or N—H. Exemplary polar aprotic solvents include acetone,N,N-dimethylformamide, acetonitrile, ethyl acetate, dichloromethane,tetrahydrofuran, and dimethyl sulfoxide.

A “non-polar solvent” as used herein means a solvent having a lowdialectric constant (<5) and low dipole moment of about 0.0 to about1.2. Exemplary nonpolar solvents include pentane, hexane, cyclohexane,benzene, toluene, chloroform, and diethyl ether.

For purposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.

Examples of alcohols include, without limitation, methanol, ethanol,2-butoxyethanol, propanol, allyl alcohol, methallyl alcohol, prenol,isopropanol, 2,2-dimethylpropan-1-ol, 2-methyl-2-phenylpropan-1-ol,butanol, isobutanol, sec-butanol, tert-butanol, 2-buten-1-ol, pentanol,2-cyclopenten-1-ol, 4-cyclopenten-1-ol, cyclopentanol,3-cyclopenten-1-ol, hexanol, cyclohexanol, 3-cyclohexen-1-ol, phenol,1-naphthol, 2-naphthol, benzyl alcohol, menthol, 1,2-ethanediol,9-fluorenylmethanol, resorcinol, meta-cresol, cinnamyl alcohol, andgeraniol.

IV. Compositions of the Invention

In certain embodiments, the invention provides a crystalline form of acompound having the structure of formula (I), described herein.

In certain embodiments, the purity of the crystalline form is 95%, 96%,97%, 98%, 99%, 99.2%, 99.4%, 99.6%, 99.8% or higher.

In yet further embodiments, the present invention provides apharmaceutical composition comprising a crystalline form of theinvention in combination with a pharmaceutically acceptable carrier.

As described in detail below, the pharmaceutical compositions describedherein may be specially formulated for administration in solid or liquidform, including those adapted for the following: (1) oraladministration, for example, drenches (aqueous or non-aqueous solutionsor suspensions), tablets, e.g., those targeted for buccal, sublingual,and systemic absorption, boluses, powders, granules, pastes forapplication to the tongue; (2) parenteral administration, for example,by subcutaneous, intramuscular, intravenous or epidural injection as,for example, a sterile solution or suspension, or sustained-releaseformulation; or (3) sublingually.

In some embodiments, the composition comprises additional agents. Forexample, the composition may comprise a nutritional agent, such as anantioxidant. Examples of pharmaceutically-acceptable antioxidantsinclude: (1) water soluble antioxidants, such as ascorbic acid, cysteinehydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfiteand the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate,butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT),lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metalchelating agents, such as citric acid, ethylenediamine tetraacetic acid(EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

The formulations of the compounds described herein may be presented inunit dosage form and may be prepared by any methods well known in theart of pharmacy. The amount of active ingredient which can be combinedwith a carrier material to produce a single dosage form will varydepending upon the host being treated and the particular mode ofadministration. The amount of active ingredient which can be combinedwith a carrier material to produce a single dosage form will generallybe that amount of the agent which produces a therapeutic effect.

In certain embodiments, a formulation described herein comprises anexcipient, including, but not limited to, cyclodextrins, liposomes,micelle forming agents, e.g., bile acids, and polymeric carriers, e.g.,polyesters and polyanhydrides; and an agent of the invention. In someembodiments, an aforementioned formulation renders orally bioavailablean agent of the invention. Methods of preparing these formulations orcompositions may include the step of bringing into association acompound of the invention with the carrier and, optionally, one or moreaccessory ingredients.

Liquid dosage forms for oral administration of the formulations providedherein include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations provided herein suitable for oral administration may be inthe form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a compound of theinvention as an active ingredient. A compound of the invention may alsobe administered as a bolus, electuary, or paste.

In solid dosage forms of the invention for oral administration (e.g.,capsules, tablets, pills, dragees, powders, granules and the like), theactive ingredient is mixed with one or more pharmaceutically-acceptablecarriers, such as sodium citrate or dicalcium phosphate, and/or any ofthe following: (1) fillers or extenders, such as starches, lactose,sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as,for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol;(4) disintegrating agents, such as agar-agar, calcium carbonate, potatoor tapioca starch, alginic acid, certain silicates, and sodiumcarbonate; (5) solution retarding agents, such as paraffin; (6)absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as, for example, cetyl alcohol, glycerolmonostearate, and non-ionic surfactants; (8) absorbents, such as kaolinand bentonite clay; (9) lubricants, such a talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof; and (10) coloring agents. In the case of capsules,tablets and pills, the pharmaceutical compositions may also comprisebuffering agents. Solid compositions of a similar type may also beemployed as fillers in soft and hard-shelled gelatin capsules using suchexcipients as lactose or milk sugars, as well as high molecular weightpolyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions described herein, such as dragees, capsules, pills andgranules, may optionally be scored or prepared with coatings and shells,such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. Compositions described herein may also beformulated for rapid release, e.g., freeze-dried. They may be sterilizedby, for example, filtration through a bacteria-retaining filter, or byincorporating sterilizing agents in the form of sterile solidcompositions which can be dissolved in sterile water, or some othersterile injectable medium immediately before use. These compositions mayalso optionally contain opacifying agents and may be of a compositionthat they release the active ingredient(s) only, or preferentially, in acertain portion of the gastrointestinal tract, optionally, in a delayedmanner. Examples of embedding compositions which can be used includepolymeric substances and waxes. The active ingredient can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-described excipients.

Pharmaceutical compositions provided herein suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically-acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containsugars, alcohols, antioxidants, buffers, bacteriostats, solutes whichrender the formulation isotonic with the blood of the intended recipientor suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

V. Methods of Treatment

In certain embodiments, the present invention provides a method ofimproving cellular health in a subject, comprising administering to thesubject a crystalline form disclosed herein.

In further embodiments, the present invention provides methods for:

improving sleep quality;

stimulating or increasing REM sleep;

treating or preventing insomnia;

treating or preventing desynchronosis; or

treating or preventing a circadian rhythm sleep disorder in a subject,comprising administering to the subject a crystalline form disclosedherein.

In certain embodiments, such a method of treatment further comprisesadministering pterostilbene.

In some embodiments, the methods treat or prevent a circadian rhythmsleep disorder. The circadian rhythm sleep disorder can be extrinsic(e.g., shift work sleep disorder, desynchornosis) or intrinsic (e.g.,advanced sleep phase disorder (ASPD), delayed sleep phase disorder(DSPD), irregular sleep-wake rhythm, and/or non-24-hour sleep-wakedisorder (i.e., hypernychthemeral syndrome)).

The subject may be male or female. In some embodiments, the subject isan adult (i.e., 18 years of age or older). The subject may be pediatric(i.e., less than 18 years of age). In some embodiments, the subject is amammal, preferably, a human.

In certain aspects, the methods and compositions provided herein relateto improving sleep health and the quality of sleep in a subject in asubject by administering to the subject (e.g., orally administering tothe subject) a composition comprising crystalline nicotinamide ribosidein the crystalline form described herein and/or pterostilbene. Sleepquality may refer to the “restfulness” of sleep (i.e., how rested anindividual feels during waking hours). Sleep quality may refer to thequantity of sleep. Good sleep quality is associated with a wide range ofpositive outcomes such as better health, less daytime sleepiness,greater well-being and better psychological functioning. In someembodiments, a subject has a score of 1, 2, or 3 on the Pittsburgh SleepQuality Index (PSQI). Further details on the PSQI may be found atBuysse, D. J., Reynolds, C. F., Monk, T. H., Berman, S. R., & Kupfer, D.J. (1989). The Pittsburgh Sleep Quality Index (PSQI): A new instrumentfor psychiatric research and practice. Psychiatry Research, 28(2),193-213, incorporated herein by reference in its entirety. In someembodiments, the subject has trouble falling asleep. In someembodiments, the subject has trouble staying asleep. In someembodiments, the subject wakes in the morning at an hour that woulddisrupt a normal sleep cycle or otherwise affect sleep quality.

In some aspects, provided herein are methods of stimulating REM sleep inneed thereof, comprising administering to the subject a crystalline formof the invention and/or pterostilbene. Rapid eye movement sleep (REMsleep) is a unique phase of sleep characterized by rapid movement of theeyes, low muscle tone throughout the body, and the propensity of thesleeper to dream vividly. In some embodiments, the compositions andmethods disclosed herein increase the total amount of time a subject isin REM sleep per sleep session (e.g., the total amount of time in REMper night). In some embodiments, the compositions and methods disclosedherein increase the amount of time a subject is in REM sleep per sleepcycle. Sleep progresses in a series of four or five more or less regularsleep cycles of non-REM and REM sleep throughout the night. The firstsleep cycle is typically around 90 minutes in length, with thesucceeding cycles averaging around 100-120 minutes, although someindividuals may have longer or shorter average cycles. Each cyclefollows the stages of non-REM sleep (stage 1-stage 2-stage 3) and then,after a period in deep stage 3 slow-wave sleep, back through the stages(stage 3-stage 2-stage 1). Then, instead of waking, the sleeper mayenter a short period of REM sleep, before going back through non-REMstages in a new cycle. As the night progresses, the time spent in deepstage 3 sleep decreases and the time spent in REM sleep increases, sothat there is a greater proportion of stage 3 sleep earlier in thenight, and a greater proportion of REM sleep later in the night,particularly during the final two sleep cycles. As used herein,stimulating REM or increasing REM may refer to increasing the time asubject stays in REM sleep per sleep cycle or the total amount of time asubject is sleeping per day.

In certain aspects, the methods and compositions provided herein relateto the treatment and/or prevention of sleep disorders in a subject byadministering to the subject (e.g., orally administering to the subject)a crystalline form of the invention and/or pterostilbene.

In some embodiments, the subject has insomnia. Insomnia is a sleepdisorder that is characterized by the inability to sleep. For example, asubject with insomnia may have trouble falling asleep, staying asleep,or wake up too early and not be able to get back to sleep. As usedherein, insomnia may refer to short-term (acute) insomnia (i.e., theinability to sleep that lasts for days to weeks) or long-term (chronic)insomnia (i.e., the inability to sleep for one month or more). In someembodiments, the insomnia is transient insomnia. Insomnia may be theresult of stress, a traumatic event, nasal/sinus allergies,gastrointestinal problems, brain lesions and tumors, stroke, chronicpain, chronic fatigue syndrome, congestive heart failure, angina,acid-reflux disease (GERD), chronic obstructive pulmonary disease,asthma, endocrine disorders such as hyperthyroidism, arthritis,neurological conditions such as Parkinson's or Alzheimer's disease, lowback pain, or genetics. In some embodiments, the subject has restlessleg syndrome. In some embodiments, the subject has sleep apnea. In someembodiments, the subject has a psychological condition that interfereswith sleep, such as anxiety, depression, bipolar disorder,schizophrenia, posttraumatic stress disorder (PTSD), and/or attentiondeficit hyperactivity disorder (ADHD). In some embodiments, insomnia isa side effect of medication. Examples of medications that may causeinsomnia include, but are not limited to, corticosteroids, alphablockers, beta blockers, SSRI antidepressants, ACE inhibitors,cholinesterase inhibitors, second generation (non-sedating) HI agonists,or glucosamine/chondroitin.

Provided herein are methods and compositions useful in regulating asubject's circadian rhythm. Circadian rhythms regulate the timing ofperiods of sleepiness and wakefulness throughout the day. Circadianrhythms are endogenously generated, although they can be modulated byexternal cues such as sunlight and temperature. Circadian rhythms areimportant in determining the sleeping and feeding patterns of allanimals, including human beings. There are clear patterns of brain waveactivity, hormone production, cell regeneration and other biologicalactivities linked to this daily cycle, and an irregular circadian rhythmmay lead to a disturbance any of the previously mentioned processes. Insome embodiments, the subject has a circadian rhythm sleep disorder. Thecircadian rhythm sleep disorder may be extrinsic (e.g., the result ofenvironmental influences or circumstances) or intrinsic (e.g., theresult of genetics or not the result of circumstances). An example of anextrinsic circadian sleep disorder includes shift work sleep disorder,which often affects individuals who work nights or in rotating shifts.Intrinsic sleep disorders include advanced sleep phase disorder (ASPD),delayed sleep phase disorder (DSPD), irregular sleep-wake rhythm, and/ornon-24-hour sleep-wake disorder (i.e., hypernychthemeral syndrome)).ASPD is characterized by difficulty staying awake in the evening anddifficulty staying asleep in the morning. DSPD is characterized by amuch later than normal timing of sleep onset and offset and a period ofpeak alertness in the middle of the night. Individuals with irregularsleep-wake rhythm suffer from sleeping at very irregular times, andusually more than twice per day (waking frequently during the night andtaking naps during the day), but often sleep a normal period of totaltime per day typical for the person's age. Non-24-hour sleep-wakedisorder, or hypernychthemeral syndrome, is a sleep disorder wherein theaffected individual's sleep occurs later and later each day, with theperiod of peak alertness also continuously moving around the clock fromday to day.

In some aspects, the compositions and methods provided herein are usefulin treating desynchronosis (i.e., jet lag). Jet lag is a temporary sleepdisorder caused by crossing time zones (e.g., during an airplaneflight), and is often the result of disruption to the circadian rhythmsof the body. Jet lag may occur any time the body's internal clock is outof sync with cues from a new time zone. Cues can include light exposureand eating times. General symptoms include fatigue and disorientation,interrupted sleep, confusion, mood changes, and pain in limbs.

Actual dosage levels and administration regimen of the compositionsdisclosed herein may be varied so as to obtain an amount of nicotinamideriboside and/or pterostilbene that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient. In some embodiments,the subject continuously self-administers the compounds disclosedherein. In other embodiments, the subject may take a compound disclosedherein as needed.

In some embodiments, administration of the composition comprisesadministration of the composition in one or more dose(s). In someembodiments, administration of the composition comprises administrationof the composition in one or more, five or more, ten or more, twenty ormore, thirty or more, forty or more, fifty or more, one hundred or more,or one thousand or more dose(s). In some embodiments, the dose comprisesat least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, atleast 125 mg, at least 150 mg, at least 200 mg, at least 225 mg, atleast 250 mg, at least 275 mg, at least 300 mg, at least 325 mg, atleast 350 mg, at least 375 mg, at least 400 mg, at least 425 mg, atleast 450 mg, at least 475 mg, at least 500 mg, at least 550 mg, atleast 600 mg, at least 650 mg, at least 700 mg, at least 750 mg, atleast 800 mg, or at least 850 mg nicotinamide riboside (compound 4). Insome embodiments, the dose comprises at least 5 mg, at least 10 mg, atleast 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, atleast 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80mg, at least 90 mg, at least 95 mg, at least 100 mg, at least 110 mg, atleast 120 mg, at least 130 mg, at least 140 mg, at least 150 mg, atleast 160 mg, least 170 mg, at least 180 mg, at least 190 mg, at least200 mg, or at least 250 mg of pterostilbene.

The compositions disclosed herein may be administered over any period oftime effective to achieve the desired therapeutic response for aparticular patient, composition, and mode of administration, withoutbeing toxic to the patient. The period of time may be at least 1 day, atleast 10 days, at least 20 days, at least 30, days, at least 60 days, atleast three months, at least six months, at least a year, at least threeyears, at least five years, or at least ten years. The dose may beadministered when needed, sporadically, or at regular intervals. Forexample, the dose may be administered monthly, weekly, biweekly,triweekly, once a day, or twice a day.

In further embodiments, the present invention provides methods of:

treating or preventing a motor neuron disease;

treating or preventing ALS; or

slowing or reversing the progression of motor neuron degeneration in asubject, comprising administering to the subject a crystalline formdisclosed herein.

In certain embodiments, such a method of treatment further comprisesadministering pterostilbene.

In certain embodiments, the motor neuron disease is ALS, hereditaryspastic paraplegia (HSP), primary lateral sclerosis (PLS), progressivemuscular atrophy (PMA), progressive bulbar palsy (PBP), pseudobulbarpalsy, or a spinal muscular atrophy.

In some embodiments, the subject may have or be predisposed to a motorneuron disease (e.g., amyotrophic lateral sclerosis (ALS), such asmedulla ALS or brainstem ALS). A motor neuron disease or disorder may beany disease or disorder that affects the function or structure of motorneuron. As used herein, a motor neuron diseases include progressivediseases that result in loss of function of motor neurons, or nerves, inthe brain and spinal cord. Examples of motor neuron diseases includeamyotrophic lateral sclerosis (ALS), hereditary spastic paraplegia(HSP), primary lateral sclerosis (PLS), progressive muscular atrophy(PMA), progressive bulbar palsy (PBP), pseudobulbar palsy, or a spinalmuscular atrophy. A motor neuron disease may affect the upper motorneurons or the lower motor neurons.

Actual dosage levels and administration regimen of the compositionsdisclosed herein may be varied so as to obtain an amount of nicotinamideriboside and/or pterostilbene that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

In some embodiments, administration of the composition comprisesadministration of the composition in one or more dose(s). In someembodiments, administration of the composition comprises administrationof the composition in one or more, five or more, ten or more, twenty ormore, thirty or more, forty or more, fifty or more, one hundred or more,or one thousand or more dose(s). In some embodiments, the dose comprisesat least 100 mg, at least 200 mg, at least 300 mg, at least 400 mg, atleast 500 mg, at least 600 mg, at least 700 mg, at least 800 mg, atleast 900 mg, at least 1000 mg, at least 1100 mg, at least 1200 mg, atleast 1300 mg, at least 1400 mg, at least 1500 mg, at least 1600 mg, atleast 1700 mg, at least 1800 mg, at least 1900 mg, at least 2000 mg, atleast 2100 mg, at least 2200 mg, at least 2300 mg, at least 2400 mg, atleast 2500 mg, at least 2600 mg, at least 2700 mg, at least 2800 mg, atleast 2900 mg, or at least 3000 mg, of nicotinamide riboside compound4). In some embodiments, the dose comprises at least 5 mg, at least 10,at least 20 mg, at least 30 mg, at least 40 mg, at least 50 mg, at least60 mg, at least 80 mg, at least 100 mg, at least 120 mg, at least 140mg, at least 160 mg, at least 180 mg, at least 200 mg, at least 220 mg,at least 240 mg, at least 260 mg, at least 280 mg, at least 300 mg, atleast 320 mg, at least 340 mg, at least 360 mg, at least 380 mg, atleast 400 mg, at least 500 mg, at least 600 mg, at least 700 mg, atleast 800 mg, at least 900 mg, or at least 1000 mg of pterostilbene.

The compositions disclosed herein may be administered over any period oftime effective to achieve the desired therapeutic response for aparticular patient, composition, and mode of administration, withoutbeing toxic to the patient. The period of time may be at least 1 day, atleast 10 days, at least 20 days, at least 30, days, at least 60 days, atleast three months, at least six months, at least a year, at least threeyears, at least five years, or at least ten years. The dose may beadministered when needed, sporadically, or at regular intervals. Forexample, the dose may be administered monthly, weekly, biweekly,triweekly, once a day, or twice a day.

In some embodiments, the subject is given a test to measure the generalprogression or symptomatic progression of a motor neuron disease. Insome embodiments, the subject is given a motor function test and/or acognition and conduct function test. The motor function test may beRevised Amyotrophic Lateral Sclerosis Functional Rating Scale(ALSFRS-R). The cognition and conduct test may be Complutense VerbalLearning Test (TAVEC), Symbol Digit Modalities Test (SDMT), VerbalFluency Test, Digit Span (Wechsler Memory Scale III), D2 Attention Test,Wechsler Memory Scale III for Letters and Numbers, London Tower Test,Stroop test, Frontal System Behavior Scale (FrSBe), and/or Brief Test(subjective conduct). In some embodiments, subjects are given both motorfunction and cognitive function tests. Motor function or cognitivefunctions tests may be given to the subject once or multiple times.

In some embodiments, the method further comprises measuring a feature(e.g., a feature associated with inflammation) in the subject. In someembodiments, the feature is measured in a blood test. Examples offeatures that may be tested are the level of a cytokine, level ofamyloid A protein, level of macrophage activation marker neopterin,level of creatine phosphokinase (CPK), level of erythrocytesedimentation rate, level of C-reactive protein, plasma viscosity,and/or white blood cell count. In some embodiments, the cytokine isproinflammatory cytokine. In some embodiments, the cytokine is ananti-inflammatory cytokine. Examples of cytokines include, but are notlimited to, TNFα, IFNγ, IL-1, IL-6, IL-8, or TGFβ.

In some embodiments, the method further comprises administering a fattyacid supplement to the subject. In some embodiments, the fatty acidsupplement comprises an oil. The oil may be processed (e.g., refined,bleached, or deodorized). In other embodiments, the oil is unprocessedor a virgin oil. In some embodiments, the fatty acid supplement isderived or fractionated from a source to yield separated fatty acids. Insome embodiments, the oil is a coconut oil. Coconut oil, as used herein,may include any oil produced by the nut of the coconut palm. Fatty acidsfound in the supplements disclosed herein may be short-chain fattyacids, medium chain fatty acids, or long chain fatty acids. Exemplaryfatty acids that may be found in the supplement include, but are notlimited to, caproic acid, caprylic acid, capric acid, lauric acid,myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleicacid, linoleic acid, and/or linolenic acid. The fatty acid supplementdisclosed herein may comprise saturated fatty acids, unsaturated fattyacids, monounsaturated fatty acids, and/or polyunsaturated fatty acids.In some embodiments, the fatty acid supplement may comprise ahydrogenated oil. Fatty acid supplements may comprise one or more fattyacid(s). Actual dosage levels and administration regimen of the fattyacid supplement disclosed herein may be varied so as to obtain an amountof fatty acid supplement that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

In further embodiments, the present invention provides methods relatingto:

improving fertility;

treating or preventing infertility;

inducing ovulation;

increasing sperm count; or

increasing lactation;

comprising administering to the subject a crystalline form of theinvention.

In certain embodiments, such a method of treatment further comprisesadministering pterostilbene.

In some embodiments, improving fertility or treating and/or preventinginfertility comprises inducing ovulation and/or oocyte and folliclematuration in a female subject. In some embodiments, improving fertilityor treating and/or preventing infertility comprising increasing spermcount and/or sperm motility in a male subject.

In some embodiments, the subject is a mammal (e.g., a human, a non-humanmammal). The subject may be male or female. In some embodiments, thesubject has impaired fertility (e.g., reduced sperm count, reduced spermmotility, reduced ovulation, reduced follicle and/or oocyte maturation).In some embodiments, the subject is infertile or sterile.

In some embodiments, provided herein are methods of increasing overallsperm health and/or sperm count in a subject by administering to thesubject (e.g., a subject in need thereof) a composition disclosedherein. Increasing sperm count may be achieved by increasing theconcentration of spermatozoa in seminal fluid, increasing the absolutenumber of spermatozoa in semen and/or increasing the volume of semen perejaculate. The methods and compositions disclosed herein may increaseoverall sperm count by increasing or inducing spermatogenesis. In someembodiments, administering the compositions disclosed herein increase orimprove sperm motility (e.g., increasing the percentage of spermatozoamoving in semen or increasing the amount of time spermatozoa are moving)in a subject. In some embodiments, administering the compositionsdisclosed herein maintains or improves overall sperm health, spermcount, and/or sperm motility in the testes and/or epididymis postspermatogenesis.

In some embodiments, provided herein are methods of inducing and/orincreasing the likelihood of ovulation in a subject by administering tothe subject (e.g., a subject in need thereof) a composition disclosedherein. In some embodiments, provided herein are methods of inducingfollicle and oocyte maturation (e.g., folliculogenesis) in a subject byadministering to the subject (e.g., a subject in need thereof) acomposition disclosed herein.

Disclosed herein are compositions and methods to aid in in vitrofertilization procedures and practices. In some aspects, provide hereinare methods of treating, preserving, or improving gamete (i.e., spermand/or oocyte) likelihood of fertilization in an in vitro procedure. Insome embodiments, compositions disclosed herein are added to semen(e.g., semen obtained from a donor subject) in preparation of forartificial insemination or intrauterine insemination (IUI). In someembodiments, compositions disclosed herein are added to semen inpreparation intracytoplasic sperm injection into an oocyte.

In some embodiments, the compositions disclosed herein may be used toenhance mitochondrial numbers, mitochondrial activity, cellular energylevels or cellular energy-producing potential in oocytes, postnatalfemale germline stem cells (also referred to herein as OSCs) and/orpreimplantation embryos prior to conducting and/or following methods ofin vitro fertilization. It has been recently discovered that adultfemale mammals retain rare germline or oogonial stem cells (OSCs) thatroutinely produce new oocytes in a manner analogous to germline stemcell support of sperm production in the adult testis, and these OSCs maybe new targets for in vitro fertilization therapies (Spradling, Nature2004 428:133-134). In some embodiments, provided herein are methods ofincreasing the overall viability of an oocyte removed from a subject(e.g., an oocyte removed in preparation for in vitro fertilization). Insome embodiments, an oocyte is treated or stored with a compositiondisclosed herein prior to in vitro fertilization. In one example,provided herein are methods of in vitro fertilization, the methodinvolving the steps of: incubating an oocyte from a subject with acomposition disclosed herein; and fertilizing the oocyte in vitro toform a zygote. In another example, the methods provided herein include amethod of in vitro fertilization, the method involving the steps of (a)incubating an OSC from a subject with composition disclosed herein; (b)obtaining a composition containing OSC mitochondria from the OSC; (c)transferring the composition into an isolated oocyte (e.g., an oocyteextracted from a subject); and (d) fertilizing the oocyte in vitro toform a zygote.

In some embodiments, the composition disclosed herein may be added to asolution used for in vitro fertilization procedures for oocytepreparation and/or storage, such as cell culture medium, oocyteretrieval solution, oocyte washing solution, oocyte in vitro maturationmedium, ovarian follicle in vitro maturation medium, oocyte in vitrofertilization medium, embryo culture medium, cleavage medium,vitrification solution, cryopreservation solution and/or embryo thawingmedium.

Gametes may be stored for any period of time with the compositionsdisclosed herein before in vitro fertilization is performed.

In some aspects, provided herein are methods of increasing lactation ina subject by administering to the subject (e.g., a subject in needthereof) a composition disclosed herein. In some embodiments, increasinglactation comprising increasing the rate at which milk is secretedand/or produced from the mammary glands of a subject. In someembodiments, increasing lactation comprises increasing the volume ofsecreted milk in the subject. In some embodiments, the subject is ahuman. In some embodiments the subject is a non-human animal, such as adairy animal (e.g., a cow, a buffalo, a goat, a sheep, a camel).

In some aspects, provided herein are methods of improving animalfecundity and/or breeding outcomes in animal husbandry by administeringthe compositions disclosed herein to a subject(s) (e.g., a non-humansubject). In some embodiments, increasing litter size in a subject(e.g., a non-human subject, such as a domesticated animal). In someembodiments, the subject is a mammal. The subject may be a rodent,lagomorph, feline, canine, porcine, ovine, bovine, equine, or primate.For example, a composition disclosed herein may be administered to afemale subject to increase number of offspring per litter orreproductive cycle. Alternately, the compositions disclosed herein maybe administered to male subjects to increase spermatozoa production toobtain semen samples with increased virility for use in artificialinsemination.

Actual dosage levels and administration regimen of the compositionsdisclosed herein may be varied so as to obtain an amount of nicotinamideriboside and/or pterostilbene that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

In some embodiments, administration of the composition comprisesadministration of the composition in one or more dose(s). In someembodiments, administration of the composition comprises administrationof the composition in one or more, five or more, ten or more, twenty ormore, thirty or more, forty or more, fifty or more, one hundred or more,or one thousand or more dose(s). In some embodiments, the dose comprisesat least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, atleast 125 mg, at least 150 mg, at least 200 mg, at least 225 mg, atleast 250 mg, at least 275 mg, at least 300 mg, at least 325 mg, atleast 350 mg, at least 375 mg, at least 400 mg, at least 425 mg, atleast 450 mg, at least 475 mg, at least 500 mg, at least 550 mg, atleast 600 mg, at least 650 mg, at least 700 mg, at least 750 mg, atleast 800 mg, or at least 850 mg of nicotinamide riboside (compound 4).In some embodiments, the dose comprises at least 5 mg, at least 10 mg,at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg,at least 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least80 mg, at least 90 mg, at least 95 mg, at least 100 mg, at least 110 mg,at least 120 mg, at least 130 mg, at least 140 mg, at least 150 mg, atleast 160 mg, least 170 mg, at least 180 mg, at least 190 mg, at least200 mg, or at least 250 mg of pterostilbene.

The compositions disclosed herein may be administered over any period oftime effective to achieve the desired therapeutic response for aparticular patient, composition, and mode of administration, withoutbeing toxic to the patient. The period of time may be at least 1 day, atleast 10 days, at least 20 days, at least 30, days, at least 60 days, atleast three months, at least six months, at least a year, at least threeyears, at least five years, or at least ten years. The dose may beadministered when needed, sporadically, or at regular intervals. Forexample, the dose may be administered monthly, weekly, biweekly,triweekly, once a day, or twice a day.

In further embodiments, the present invention provides methods relatingto:

treating or preventing kidney damage;

treating or preventing acute kidney injury;

treating or preventing kidney disease; or

increasing blood flow to the kidneys;

comprising administering to the subject a crystalline form of theinvention.

In certain embodiments, such a method of treatment further comprisesadministering pterostilbene.

In certain embodiments, the kidney damage is the result of cancer,decreased blood flood to the kidneys (i.e., ischemia), back up of urinein the kidneys, sepsis, trauma, autoimmune disease, drug-inducedtoxicity (e.g., non-steroidal anti-inflammatory (MAID) inducednephrotoxicity), lead poisoning, and/or severe dehydration.

In some embodiments, the acute kidney injury is the result of decreasedblood flow to the kidneys (i.e., ischemic injury). Decreased blood flowto the kidneys may be a result of hypotension, blood loss, severediarrhea, heat attack, heart failure, deceased heart function, organfailure, drug-induced nephrotoxicity (e.g., NSAID inducednephrotoxicity), allergic reactions, burns, trauma (e.g., blunt trauma),and/or surgery. In some embodiments, the acute kidney injury is theresult of cancer (e.g., multiple myeloma), sepsis, vasculitis,interstitial nephritis, scleroderma, tubular necrosis,glomerulonephritis, or thrombotic microangiopathy. In some embodiments,the acute kidney injury is the result of blockage of the urinary tract.Blockage of the urinary tract may be caused by bladder cancer, prostatecancer, cervical cancer, an enlarged prostate, kidney stones, or bloodclots in the urine.

In some embodiments, the chronic kidney disease is the result of animmune system disease (e.g., lupus), long term viral disease (e.g.,HIV/AIDS, hepatitis B, or hepatitis C), urinary tract infections,polycystic kidney disease, and/or inflammation of glomeruli.

As used herein, kidney damage may refer to a medical condition ofimpaired kidney function in which the kidneys fail to adequately filtermetabolic wastes from the blood. In some embodiments, kidney damage isalso indicative of kidney failure. Examples of conditions that may causekidney damage include, but are not limited to decreased blood flood tothe kidneys, back up of urine in the kidneys, sepsis, trauma (e.g., suchas blunt trauma), an autoimmune disease, drug-induced nephrotoxicity(e.g., NSAID induced nephrotoxicity), heavy mental poisoning (e.g., leadpoisoning), or severe dehydration.

In some aspects, provided herein are methods of treating or preventingacute kidney injury (i.e., in a subject in need thereof). In someembodiments, acute kidney injury is an episode of kidney failure orkidney damage that happens within a few hours or a few days. Acutekidney injury, as used herein, may be characterized by abruptdeterioration in kidney function. In some embodiments, the subject hasacute kidney injury, and the acute kidney injury may manifest by anincrease in serum creatinine level with or without reduced urine output.In some embodiments, the subject has increased serum creatinine levels.In some embodiments, the subject may have reduced urine output. In someembodiments, the subject has acute kidney injury, and the acute kidneyinjury may be prerenal (e.g., caused by decreased renal blood flow),intrinsic renal (e.g., caused by a process within the kidneys), orpostrenal (e.g., caused by inadequate drainage of urine distal to thekidneys). Acute kidney injury or kidney damage may be a result of use(e.g., overuse) of medications, such as NSAIDs, angiotensin-convertingenzyme inhibitors, angiotensin receptor blockers, cyclosporine,diuretics, tacrolimus, penicillin analogues, cephalosporins,sulfonamides, ciprofloxacin, acyclovir, rifampin, phenytoin, interferon,or proton pump inhibitors. Other causes of acute kidney injury andkidney damage include cardiorenal syndrome, hepatorenal syndrome,abdominal compartment syndrome, hypercalcemia, sepsis, neurogenic shock,infections of the renal parenchyma, glomerulonephritis, viral infections(such as Epstein-Barr virus infections or cytomegalovirus infections),bacterial infections (e.g., bacterial infections caused by bacteria ofthe Streptococcus or Legionella species), or fungal infections (e.g.,fungal infections caused by candidiasis or histoplasmosis). In someembodiments, the acute renal injury and/or kidney damage is caused by asystemic disease, such as sarcoidosis or lupus.

Additional examples of conditions that cause acute kidney injury andkidney damage include cancer (e.g., multiple myeloma), prolongedhypotension, renal vein thrombosis, malignant hypertension, sclerodermarenal crisis, renal atheroembolic disease, renal infarction vasculitis,interstitial nephritis, scleroderma, and or conditions that causeinflammation of or damage to the kidney tubules, such as tubularnecrosis, glomerulonephritis, or thrombotic microangiopathy.

In some embodiments, the acute kidney injury and/or kidney damage iscaused by a decrease in blood flow to the kidney. Conditions that maycause a decrease in blood flow to the kidneys includes, for example,blood loss, severe diarrhea, heat attack, heart failure, deceased heartfunction, organ failure, allergic reactions, burns, and/or trauma. Insome embodiments, the subject has undergone surgery, and the subject'sblood vessels have been clamped, leading to a decrease of blood flow tothe kidneys. In some embodiments, the acute kidney injury or kidneydamage is the result of a blockage of the urinary tract. A blockage ofthe urinary tract may be the result of, for example, neurogenic bladder,retroperitoneal fibrosis, bladder cancer, prostate cancer, cervicalcancer, an enlarged prostate, kidney stones, blood clots, or tumors.

In some embodiments, the subject has a kidney disease. A kidney diseaseis any condition that affects the kidney's ability to filter compoundsout of blood, filter extra water out of blood, and/or help control bloodpressure. Kidney disease may be caused by diabetes, hypertension, asystemic disease (e.g., lupus), viral disease (e.g., HIV/AIDS, hepatitisB, or hepatitis C), urinary tract infections, a genetic disease, such aspolycystic kidney disease, or any condition that results in theinflammation of kidney glomeruli.

In some embodiments, the subject's kidney function may be measuredprior, during or after administration of a composition disclosed herein.Kidney function may be evaluated as a function of glomerular filtrationrate, urine output, or the level of other biomedical markers of kidneyhealth, such as creatinine, urea, nitrogen, phosphorus, or potassium.Markers such as creatinine, urea, nitrogen, phosphorus, or potassium maybe measured in the urine or through a blood test.

Actual dosage levels and administration regimen of the compositionsdisclosed herein may be varied so as to obtain an amount of nicotinamideriboside and/or pterostilbene that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

In some embodiments, administration of the composition comprisesadministration of the composition in one or more dose(s). In someembodiments, administration of the composition comprises administrationof the composition in one or more, five or more, ten or more, twenty ormore, thirty or more, forty or more, fifty or more, one hundred or more,or one thousand or more dose(s). In some embodiments, the dose comprisesat least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, atleast 125 mg, at least 150 mg, at least 200 mg, at least 225 mg, atleast 250 mg, at least 275 mg, at least 300 mg, at least 325 mg, atleast 350 mg, at least 375 mg, at least 400 mg, at least 425 mg, atleast 450 mg, at least 475 mg, at least 500 mg, at least 550 mg, atleast 600 mg, at least 650 mg, at least 700 mg, at least 750 mg, atleast 800 mg, or at least 850 mg of nicotinamide riboside (Compound 4).In some embodiments, the dose comprises at least 5 mg, at least 10 mg,at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg,at least 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least80 mg, at least 90 mg, at least 95 mg, at least 100 mg, at least 110 mg,at least 120 mg, at least 130 mg, at least 140 mg, at least 150 mg, atleast 160 mg, least 170 mg, at least 180 mg, at least 190 mg, at least200 mg, or at least 250 mg of pterostilbene. The compositions disclosedherein may be administered over any period of time effective to achievethe desired therapeutic response for a particular patient, composition,and mode of administration, without being toxic to the patient. Theperiod of time may be at least 1 day, at least 10 days, at least 20days, at least 30, days, at least 60 days, at least three months, atleast six months, at least a year, at least three years, at least fiveyears, or at least ten years. The dose may be administered when needed,sporadically, or at regular intervals. For example, the dose may beadministered monthly, weekly, biweekly, triweekly, once a day, or twicea day.

In further embodiments, the present invention provides methods relatingto treating and/or preventing liver related diseases and disorders andfor improving liver health in a subject by administering to the subjecta crystalline form disclosed herein. Specifically, the inventionprovides a method of:

treating or preventing liver damage;

treating or preventing fatty liver;

decreasing the serum level of alanine transaminase (ALT); or decreasingthe serum level of aspartate transaminase (AST) in a subject;

comprising administering to the subject a crystalline form discloseddisclosed herein.

In certain embodiments, such a method of treatment further comprisesadministering pterostilbene.

In certain embodiments, the liver damage is the result of cancer (e.g.,liver cancer, bile duct cancer and/or a liver adenoma), cirrhosis, viralinfection (e.g., hepatitis A infection, a hepatitis B infection and/or ahepatitis E infection), congenital disorders of metabolism, trauma,autoimmune disease (e.g., autoimmune hepatitis, primary biliarycirrhosis, or primary sclerosing cholangitis), hemochromatosis,hyperoxaluria, oxalosis, Wilson's disease and/or drug-inducedhepatotoxicity (e.g., alcohol-induced hepatotoxicity and/oracetaminophen-induced hepatotoxicity).

Provided herein are methods of preventing or treating liver damageand/or fatty liver in a subject by administering to the subject (e.g., asubject in need thereof) a composition disclosed herein.

In some embodiments, the subject may have or be predisposed to liverdamage and/or fatty liver. Liver damage may result from any conditionthat causes the cells of the liver (i.e., hepatocytes) to die orotherwise not function normally. Examples of conditions that may causeliver damage include, but are not limited to, cancer (e.g., livercancer, bile duct cancer, or a liver adenoma), trauma, congenitalmetabolic disorders (e.g., genetic metabolic disorders resulting in anenzyme deficiency), vascular injury, cirrhosis, a viral infection (e.g.,hepatitis A, hepatitis B, hepatitis E), an autoimmune disease (e.g.,autoimmune hepatitis, primary biliary cirrhosis, or primary sclerosingcholangitis), hemochromatosis, hyperoxaluria, oxalosis, Wilson'sdisease, or drug-induced hepatotoxicity (e.g., alcohol-inducedhepatotoxicity or acetaminophen-induced hepatotoxicity). Fatty liver maybe caused by any condition that causes fat accumulation of liver. Theseconditions may be, but are not limited to, non-alcoholic fatty liverdisease or alcoholic liver disease.

Disclosed herein are methods of treating or preventing age-relatedsymptoms or diseases comprising administering a composition disclosedherein. Provided herein are methods of decreasing the amount of alaninetransaminase (ALT) and/or aspartate transaminase (AST) in a subjectcomprising administering to the subject a composition provided herein.AST and ALT are reasonably sensitive indicators of liver damage orinjury from different types of diseases or conditions, and they areoften measured in liver tests or liver blood tests. Elevated levels ofAST and ALT are associated with liver damage and liver malfunction. Insome embodiments, ALT is decreased in the subject by at least 0.1 U/L,at least 0.2 U/L, at least 0.3 U/L, at least 0.4 U/L, at least 0.5 U/L,at least 0.6 U/L, at least 0.7 U/L, at least 0.8 U/L, at least 0.9 U/L,at least 01.0 U/L, 1.1 U/L, at least 1.2 U/L, at least 1.3 U/L, at least1.4 U/L, at least 1.5 U/L, at least 1.6 U/L, at least 1.7 U/L, at least1.8 U/L, at least 1.9 U/L, at least 2.0 U/L, 2.1 U/L, at least 2.2 U/L,at least 2.3 U/L, at least 2.4 U/L, at least 2.5 U/L, at least 2.6 U/L,at least 2.7 U/L, at least 2.8 U/L, at least 2.9 U/L, at least 3.0 U/L,at least 3.5 U/L, 4.0 U/L, at least 4.5 U/L, or at least 5.0 U/L afteradministration of the composition. In some embodiments, the ALT isdecreased by at least 0.1 U/L, at least 0.2 U/L, at least 0.3 U/L, atleast 0.4 U/L, at least 0.5 U/L, at least 0.6 U/L, at least 0.7 U/L, atleast 0.8 U/L, at least 0.9 U/L, at least 01.0 U/L, 1.1 U/L, at least1.2 U/L, at least 1.3 U/L, at least 1.4 U/L, at least 1.5 U/L, at least1.6 U/L, at least 1.7 U/L, at least 1.8 U/L, at least 1.9 U/L, at least2.0 U/L, 2.1 U/L, at least 2.2 U/L, at least 2.3 U/L, at least 2.4 U/L,at least 2.5 U/L, at least 2.6 U/L, at least 2.7 U/L, at least 2.8 U/L,at least 2.9 U/L, at least 3.0 U/L, at least 3.5 U/L, 4.0 U/L, at least4.5 U/L, or at least 5.0 U/L after administration of the composition.

Nicotinamide adenine dinucleotide (NAD⁺) is a coenzyme that participatesin many metabolic reactions. NAD+ plays an important role intranscription regulation, longevity, and age-associated diseases. NAD+levels decrease with age, while increased NAD+ levels are associatedwith robust health. In some embodiments, provided herein are methods ofincreasing the amount of NAD+ of a subject by administering acomposition disclosed herein. NAD+ may increase by at least 1.0 μg/mL,at least 2.0 μg/mL, at least 3.0 μg/mL, at least 4.0 μg/mL, at least 5.0μg/mL, at least 6.0 μg/mL, at least 7.0 μg/mL, at least 8.0 μg/mL, atleast 9.0 μg/mL, at least 10.0 μg/mL, at least 11.0 μg/mL, at least 12.0μg/mL, at least 13.0 μg/mL, at least 14.0 μg/mL, at least 15.0 μg/mL, atleast 16 μg/mL, at least 17 μg/mL, at least 18 μg/mL, at least 19 μg/mL,at least 20 μg/mL, at least 21 μg/mL, at least 22 μg/mL, at least 23μg/mL, at least 24 μg/mL, at least 25 μg/mL, at least 26 μg/mL, at least27 μg/mL, at least 28 μg/mL, at least 29 μg/mL, or at least 30 μg/mLafter administration of the composition.

Provided herein are methods of decreasing blood pressure (e.g.,diastolic blood pressure) of a subject by administering a compositionherein. In some embodiments, the subject's diastolic blood pressuredecreases by at least 1 mmHg, at least 1.5 mmHg, at least 2 mmHg, atleast 2.5 mmHg, at least 3 mmHg, at least 3.5 mmHg, at least 4.0 mmHg,at least 4.5 mmHg, or at least 5 mmHg after administration of thecomposition.

In certain embodiments, the invention also provides a method of treatinga disease comprising administering to a subject a crystalline form ofthe invention.

In certain such embodiments, the disease is a neurodegenerative disease.Exemplary neurodegenerative diseases include Alzheimer's, Parkinson's,and Huntington's Disease.

In alternative such embodiments, the disease is a skin disorder. Skindisorders may be caused by exposure to the sun; exemplary such disordersare selected from the group consisting of actinic keratoses, lentiginesor age spots, seborrheic keratoses, sun burn, photosensitivity, moles,polymorphous light eruption, solar elastosis or wrinkles, skin cancer(such as melanoma, squamous cell carcinoma, and basal cell carcinoma),and freckles. Skin disorders may also be caused by inflammation;exemplary such disorders are selected from the group consisting ofpsoriasis, contact dermatitis, atopic dermatitis, seborrheic dermatitis,asteatotic eczema, discoid eczema, hand eczema, gravitational/varicoseeczema, eczematous drug eruptions, lichen simplex, acne, lichen planus,pityriasis lichenoides, keratosis lichenoides chronica, lichen nitidus,lichen striatus, mycosis fungoides, erythroderma, erythema multiforme,Stevens-Johnson Syndrome, vasculitis, and toxic epidermal necrolysis. Infurther embodiments, a skin disorder may be caused by autoimmune diseaseis selected from the group consisting of pyoderma gangrenosum, systemiclupus erythematosus, eosinophilic fasciitis, scleroderma, pemphigusvulgaris, bullous pemphigoid, alopecia areata, vitiligo, psoriasis,dermatomyositis, and dystrophic epidermolysis bullosa.

In yet alternative embodiments, the disease is obesity or diabetes. Themethods may also enable lipid lowering and blood pressure reduction.

In further alternative embodiments, the disease is muscle wasting orsarcopenia.

In further alternative embodiments, the disease is an autoimmune diseasesuch as arthritis or lupus.

In further alternative embodiments, the disease is mitochondrialdysfunction or disease.

In still further alternative embodiments, the disease is an acceleratedaging disease such as progeria.

In other embodiments, the disease is chemotherapy induced neuropathy, orcognitive decline. The methods may also enable pain reduction.

In yet further embodiments, the disease is a cancer such as breastcancer, colon cancer, etc.

In other embodiments, the invention provides methods that promotesirtuin activation, NAD+ boosting, PARP activation, autophagy andmitophagy, or mitochondrial biogenesis.

In still further embodiments, the invention provides methods ofpromoting wellness comprising administering to a subject the crystallineform of the invention.

In some embodiments, the methods promote wellness by promoting stem cellhealth and function; such stem cells may be intestinal, skin, muscle,hematopoietic, or neural stem cells.

The methods may also promote wellness by preventing hearing loss or hairloss.

Alternatives, the methods may promote healthy hair and nail growth.

In yet further alternative embodiments, the methods may promote musclebuilding for endurance and strength, weight loss, or and increasedimmune response (innate and adaptive function).

Actual dosage levels and administration regimen of the compositionsdisclosed herein may be varied so as to obtain an amount of nicotinamideriboside (or nicotinamide riboside chloride) and/or pterostilbene thatis effective to achieve the desired therapeutic response for aparticular patient, composition, and mode of administration, withoutbeing toxic to the patient.

In some embodiments, administration of the composition comprisesadministration of the composition in one or more dose(s). In someembodiments, administration of the composition comprises administrationof the composition in one or more, five or more, ten or more, twenty ormore, thirty or more, forty or more, fifty or more, one hundred or more,or one thousand or more dose(s). In some embodiments, the dose comprisesat least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, atleast 125 mg, at least 150 mg, at least 200 mg, at least 225 mg, atleast 250 mg, at least 275 mg, at least 300 mg, at least 325 mg, atleast 350 mg, at least 375 mg, at least 400 mg, at least 425 mg, atleast 450 mg, at least 475 mg, at least 500 mg, at least 550 mg, atleast 600 mg, at least 650 mg, at least 700 mg, at least 750 mg, atleast 800 mg, or at least 850 mg of nicotinamide riboside. In someembodiments, the dose comprises at least 5 mg, at least 10 mg, at least15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg,at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, at least60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg,at least 90 mg, at least 95 mg, at least 100 mg, at least 110 mg, atleast 120 mg, at least 130 mg, at least 140 mg, at least 150 mg, atleast 160 mg, least 170 mg, at least 180 mg, at least 190 mg, at least200 mg, or at least 250 mg of pterostilbene.

The compositions disclosed herein may be administered over any period oftime effective to achieve the desired therapeutic response for aparticular patient, composition, and mode of administration, withoutbeing toxic to the patient. The period of time may be at least 1 day, atleast 10 days, at least 20 days, at least 30, days, at least 60 days, atleast three months, at least six months, at least a year, at least threeyears, at least five years, or at least ten years. The dose may beadministered when needed, sporadically, or at regular intervals. Forexample, the dose may be administered monthly, weekly, biweekly,triweekly, once a day, or twice a day.

In any of the treatment methods described herein, nicotinamide ribosidechloride, alone or in combination with pterostilbene, may be formulatedtogether with one or more pharmaceutically acceptable carriers(additives) and/or diluents. In another aspect, the agents describedherein can be administered as such, or administered in mixtures withpharmaceutically acceptable carriers and can also be administered inconjunction with other agents. Conjunctive therapy thus includessequential, simultaneous and separate, or co-administration of one ormore compounds of the invention, wherein the therapeutic effects of thefirst administered has not entirely disappeared when the subsequentcompound is administered.

Compositions useful for the methods of the invention are describedabove.

EXEMPLIFICATION

The invention described generally herein will be more readily understoodby reference to the following examples, which are included merely forpurposes of illustration of certain aspects and embodiments of thepresent invention, and are not intended to limit the invention.

Example 1. Scale-Up Synthesis and Crystallization of NicotinamideRiboside Chloride

900 kg of nicotinamide riboside triacetate and 2133 kg of methanol werecharged to a reactor and mixed, then cooled to 0° C. 747 kg of 7Mmmmonia in methanol (i.e., “methanolic NH₃”) was slowly charged to thereactor at 0° C. The reaction mixture was passed through a polishfilter, then the reaction mixture was stirred for 14 hours. A samplefrom the reaction mixture was taken to assess reaction progress. Uponcompletion of the reaction, the reaction mixture was placed undervacuum, then warmed to 20° C. to 25° C. for 4 hours. Vacuum was applieduntil solids formed. Once solids were formed, the resultant slurry wasfiltered on a Nutsche filter dryer. Solids were washed with 1422 kg ofethanol, then 1422 kg of acetone, then 1322 kg of methyl tert butylether (MTBE). The resultant solids were then dried at 40° C. Product wasformed with 60% yield. The process flow diagram for this reaction isshown in FIG. 6.

Example 2. Optional Secondary Isolation

The crystalline form may optionally undergo a second isolation processaccording to the following steps: The solids obtained in Example 1 weredissolved in purified water at 30° C. to 40° C. Ethanol was slowly addedto the solution and mixed for 10 hours, over which time the solids beganto precipitate. MTBE was then added and mixed for 2 hours. The mixturewas then filtered on a Buchner funnel, and the solids were washed withethanol, then acetone, then MTBE. Solids were dried at 40° C.

Example 3. Spectroscopic Data

The crystalline form made by the process described in Examples 1 and 2has an XRD spectrum substantially as shown in FIG. 1. The instrumentutilized in collecting the XRD data is a Rigaku Smart Lab X-Raydiffraction system.

Specifically, in order to collect the XRD data, The Rigaku Smart-LabX-ray diffraction system was configured for reflection Bragg-Brentanogeometry using a line source X-ray beam. The X-ray source is a Cu LongFine Focus tube that was operated at 40 kV and 44 mA. That sourceprovides an incident beam profile at the sample that changes from anarrow line at high angles to a broad rectangle at low angles. Beamconditioning slits are used on the line X-ray source to ensure that themaximum beam size is less than 10 mm both along the line and normal tothe line. The Bragg-Brentano geometry is a para-focusing geometrycontrolled by passive divergence and receiving slits with the sampleitself acting as the focusing component for the optics. The inherentresolution of Bragg-Brentano geometry is governed in part by thediffractometer radius and the width of the receiving slit used.Typically, the Rigaku Smart-Lab is operated to give peak widths of 0.1°2θ or less. The axial divergence of the X-ray beam is controlled by5.0-degree Soller slits in both the incident and diffracted beam paths.

The samples were prepared in a low background Si holder using lightmanual pressure to keep the sample surface flat and level with thereference surface of the sample holder. The single crystal Si lowbackground holder has a small circular recess (10 mm diameter and about0.2 mm depth) that held between 20 and 25 mg of the sample. The sampleswere analyzed from 2 to 40 °2θ using a continuous scan of 6 °2θ perminute with an effective step size of 0.02 °2θ. The data collectionprocedure used to analyze these samples was not validated. The peaklists were generated using PDXL2 v.2.3.1.0. The figures were createdusing PlotMon V1.00.

INCORPORATION BY REFERENCE

All publications and patents mentioned herein are hereby incorporated byreference in their entirety as if each individual publication or patentwas specifically and individually indicated to be incorporated byreference. In case of conflict, the present application, including anydefinitions herein, will control.

EQUIVALENTS

While specific embodiments of the subject invention have been discussed,the above specification is illustrative and not restrictive. Manyvariations of the invention will become apparent to those skilled in theart upon review of this specification and the claims below. The fullscope of the invention should be determined by reference to the claims,along with their full scope of equivalents, and the specification, alongwith such variations.

1. A crystalline form of a compound having the structure of formula (I):

wherein the crystalline form is characterized by 2θ values of 20.9±0.1,21.6±0.1, 21.8±0.1, 23.6±0.1, and 24.4±0.1.
 2. The crystalline form ofclaim 1, characterized by 2θ values of 9.9±0.1, 18.4±0.1, 20.9±0.1,21.6±0.1, 21.8±0.1, 23.6±0.1, 24.4±0.1, 29.4±0.1, 29.9±0.1, 30.5±0.1,and 33.5±0.1.
 3. The crystalline form of claim 2, characterized by 2θvalues of 9.9±0.1, 18.4±0.1, 20.9±0.1, 21.6±0.1, 21.8±0.1, 22.9±0.1,23.6±0.1, 24.4±0.1, 25.2±0.1, 29.4±0.1, 29.9±0.1, 30.5±0.1, 31.9±0.1,32.1±0.1, 33.5±0.1, 34.1±0.1, and 37.4±0.1.
 4. The crystalline form ofclaim 3, characterized by 2θ values of 9.9±0.1, 15.6±0.1, 18.4±0.1,18.5±0.1, 19.0±0.1, 20.9±0.1, 21.6±0.1, 21.8±0.1, 22.9±0.1, 23.6±0.1,24.4±0.1, 25.2±0.1, 29.4±0.1, 29.9±0.1, 30.5±0.1, 31.5±0.1, 31.9±0.1,32.1±0.1, 33.5±0.1, 34.0±0.1, 34.1±0.1, 36.5±0.1, and 37.4±0.1.
 5. Thecrystalline form of claim 1, characterized by 2θ values of 20.87±0.10,21.55±0.10, 21.79±0.10, 23.63±0.10, and 24.44±0.10.
 6. The crystallineform of claim 5, characterized by 2θ values of 9.87±0.10, 18.36±0.10,20.87±0.10, 21.55±0.10, 21.79±0.10, 23.63±0.10, 24.44±0.10, 29.35±0.10,29.93±0.10, 30.47±0.10, and 33.51±0.10.
 7. The crystalline form of claim6, characterized by 2θ values of 9.87±0.10, 18.36±0.10, 20.87±0.10,21.55±0.10, 21.79±0.10, 22.87±0.10, 23.63±0.10, 24.44±0.10, 25.25±0.10,29.35±0.10, 29.93±0.10, 30.47±0.10, 31.87±0.10, 32.08±0.10, 33.51±0.10,34.15±0.10, and 37.38±0.10.
 8. The crystalline form of claim 7,characterized by 2θ values 9.87±0.10, 15.61±0.10, 18.36±0.10,18.49±0.10, 19.01±0.10, 20.87±0.10, 21.55±0.10, 21.79±0.10, 22.87±0.10,23.63±0.10, 24.44±0.10, 25.25±0.10, 29.35±0.10, 29.93±0.10, 30.47±0.10,31.46±0.10, 31.87±0.10, 32.08±0.10, 33.51±0.10, 34.00±0.10, 34.15±0.10,36.53±0.10, and 37.38±0.10.
 9. The crystalline form of claim 1, havingan XRD spectrum substantially as shown in FIG.
 1. 10. The crystallineform of claim 1, having an endotherm of 123.9° C. in the DSC thermogramat a heating rate of 5° C./min.
 11. (canceled)
 12. The crystalline formof claim 1, having an endotherm of 115.4° C. in the DSC thermogram at aheating rate of 2° C./min.
 13. (canceled)
 14. The crystalline form ofclaim 1, having an endotherm of 131.2° C. in the DSC thermogram at aheating rate of 10° C./min.
 15. (canceled)
 16. The crystalline form ofclaim 1, having an endotherm of 139.2° C. in the DSC thermogram at aheating rate of 20° C./min.
 17. (canceled)
 18. The crystalline form ofclaim 1, wherein the purity of the crystalline form is 95%, 96%, 97%,98%, 99%, 99.2%, 99.4%, 99.6%, 99.8% or higher.
 19. A pharmaceuticalcomposition, comprising a crystalline form of claim 1, and apharmaceutically acceptable carrier.
 20. A method for preparing acrystalline form of claim 1, comprising: (a) providing a mixture of acompound of formula (I) in a first organic solvent; and (b)crystallizing the compound of formula (I) from the mixture of a compoundof formula (I) in a first organic solvent. 21-30. (canceled)
 31. Amethod of improving cellular health in a subject, comprisingadministering to the subject a therapeutically effective amount of acrystalline form of claim
 1. 32. A method of improving sleep quality,stimulating or increasing REM sleep, or treating or preventing insomnia,desynchronosis, or a circadian rhythm sleep disorder in a subject,comprising administering to the subject a therapeutically effectiveamount of a crystalline form of claim
 1. 33. A method of treating orpreventing a motor neuron disease or ALS, or slowing or reversing theprogression of motor neuron degeneration in a subject, comprisingadministering to the subject a therapeutically effective amount of acrystalline form of claim
 1. 34. A method of improving fertility,treating or preventing infertility, inducing ovulation, increasing spermcount, or increasing lactation, comprising administering to the subjecta therapeutically effective amount of a crystalline form of claim
 1. 35.A method of treating or preventing kidney damage, acute kidney injury,or kidney disease, or increasing blood flow to the kidneys, comprisingadministering to the subject a therapeutically effective amount of acrystalline form of claim
 1. 36. A method of treating or preventingliver damage or fatty liver, or decreasing the serum level of alaninetransaminase (ALT) or aspartate transaminase (AST) in a subject,comprising administering to the subject a therapeutically effectiveamount of a crystalline form of claim 1.